Probe set filtering increases correlation between Affymetrix GeneChip and qRT-PCR expression measurements.
Probe set filtering increases correlation between Affymetrix GeneChip and qRT-PCR expression measurements.
复制标题
DOI:
10.1186/1471-2105-11-104
复制
发表时间:
2010-02-24
影响因子:
3
通讯作者:
Pokarowski P
中科院分区:
文献类型:
--
作者:
Mieczkowski J;Tyburczy ME;Dabrowski M;Pokarowski P
Affymetrix GeneChip microarrays are popular platforms for expression profiling in two types of studies: detection of differential expression computed by p-values of t-test and estimation of fold change between analyzed groups. There are many different preprocessing algorithms for summarizing Affymetrix data. The main goal of these methods is to remove effects of non-specific hybridization, and to optimally combine information from multiple probes annotated to the same transcript. The methods are benchmarked by comparison with reference methods, such as quantitative reverse-transcription PCR (qRT-PCR). We present a comprehensive analysis of agreement between Affymetrix GeneChip and qRT-PCR results. We analyzed the influence of filtering by fraction Present calls introduced by J.N. McClintick and H.J. Edenberg (2006) and 2 mapping procedures: updated probe sets definitions proposed by Dai et al. (2005) and our "naive mapping" method. Because of evolution of genome sequence annotations since the time when microarrays were designed, we also studied the effect of the annotation release date. These comparisons were prepared for 6 popular preprocessing algorithms (MAS5, PLIER, RMA, GC-RMA, MBEI, and MBEImm) in the 2 above-mentioned types of studies. We used data sets from 6 independent biological experiments. As a measure of reproducibility of microarray and qRT-PCR values, we used linear and rank correlation coefficients. We show that filtering by fraction Present calls increased correlations for all 6 preprocessing algorithms. We observed the difference in performance of PM-MM and PM-only methods: using MM probes increased correlations in fold change studies, but PM-only methods proved to perform better in detection of differential expression. We recommend using GC-RMA for detection of differential expression and PLIER for estimation of fold change. The use of the more recent annotation improves the results in both types of studies, encouraging re-analysis of old data.
登录
查看更多内容
DOI:
10.1073/pnas.011404098
发表时间:
2001-01-02
影响因子:
11.1
作者:
Li, C;Wong, WH
通讯作者:
Wong, WH
影响因子:
3
作者:
Sandberg R;Larsson O
通讯作者:
Larsson O
影响因子:
3
作者:
Millenaar, FF;Okyere, J;May, ST;van Zanten, M;Voesenek, LACJ;Peeters, AJM
通讯作者:
Peeters, AJM
影响因子:
3
作者:
McClintick, JN;Edenberg, HJ
通讯作者:
Edenberg, HJ
影响因子:
12.3
作者:
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者:
Zhang J